1. Field of the Invention
The present application relates generally to the treatment of neoplastic disease and the compositions employed therein. The instant invention concerns a novel phytochemotherapeutic composition, methods for producing the composition, and use of the composition in a method for treating cancer and other neoplastic diseases. Specifically, the instant invention provides for an aqueous extract, and fractions thereof obtained by chromatographic separation, isolated from Vernonia amygdalina (V. amydalina) each of which inhibit the growth of cancer cells.
2. Technical Problem Addressed
Breast cancer is the most common form of malignant disease among women in Western countries and, in the United States, is the most common cause of death among women between 40 and 55 years of age (Forrest, 1990). There is a need for improved compositions and protocols for the treatment of human breast cancer and other neoplastic diseases. The instant invention provides phytochemotherapeutic compositions and methods for inhibiting the growth of cancer cells, and specifically for the growth inhibition of human breast cancer cells.
3. Description of Related Art
Vernonia amygdalina, a plant known for its very bitter taste, has been shown to sometimes be ingested by chimpanzees suffering from parasite-related diseases (Koshimizu, 1994).
Previous investigation into the possible antineoplastic properties exhibited by Vernonia amygdalina have focused on chloroform extractable components. For example (Kupchan et al., 1969) suggests that Vernonia amygdalina contains at least three distinct cytotoxic components, namely vernolide, vernodalin, and vernomygdin (see page 3909, Chart I). However, there are no reports of any water-extractable Vernonia amygdalina components which exhibit antineoplastic properties.
The present invention provides for an anti-neoplastic pharmaceutical composition typically comprising filtrates prepared from aqueous extracts of Vernonia amygdalina leaves, collected from plants in Benin City, Nigeria. Also provided are pharmaceutical compositions prepared from fractions of the Vernonia amygdalina aqueous extracts, prepared by chromatographic separation of the extracts. The instant invention also provides for methods of preparing the Vernonia amygdalina aqueous extracts and chromatographic fractions. Additionally the instant invention provides methods for the treatment of neoplastic disease using these Vernonia amygdalina extracts and chromatographic fractions. These methods are useful for treating neoplastic disease, especially breast cancer, in animals suffering therefrom.
One embodiment of the instant invention provides for a method of preparing a pharmaceutical composition. In one aspect of this embodiment of the invention the pharmaceutical composition is produced by a process comprising the following steps:
a) Providing Vernonia amygdalina leaves.
b) Rinsing the leaves with water.
c) Soaking the leaves in water.
d) Crushing the leaves, preferably in by a gentle means, to produce a mixture.
e) Producing a filtrate by filtering the mixture to remove particulate matter.
f) Optionally, concentrating the filtrate by removing water.
Other embodiments of the instant invention provide for chromatographic separation of the aqueous extract filtrate. This separation may be by any suitable means commonly used, by those of skill in the art, to partition the various components of a complex solution and/or mixture.
Other embodiments of the instant invention provide for anti-neoplastic pharmaceutical compositions prepared from a extract, filtrate, or fraction thereof, isolated by any of the methods described above.
Other embodiments of the invention provide methods of inhibiting the growth and/or proliferation of neoplastic cells, both malignant and non-malignant. According to one aspect of this embodiment of the invention, cancer cell proliferation is inhibited by contacting the neoplastic cells with an effective amount of a pharmaceutical composition produced from an aqueous extraction of Vernonia amygdalina leaves. In another aspect of this embodiment of the invention, neoplastic cell proliferation is inhibited by contacting the cells with an effective amount of a pharmaceutical composition produced from a fraction of an aqueous extraction of Vernonia amygdalina leaves, wherein the fraction was isolated by chromatographic separation of the aqueous extract.